+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Sequence analysis of the gene encoding amylosucrase from Neisseria polysaccharea and characterization of the recombinant enzyme



Sequence analysis of the gene encoding amylosucrase from Neisseria polysaccharea and characterization of the recombinant enzyme



Journal of Bacteriology 181(2): 375-381



The Neisseria polysaccharea gene encoding amylosucrase was subcloned and expressed in Escherichia coli. Sequencing revealed that the deduced amino acid sequence differs significantly from that previously published. Comparison of the sequence with that of enzymes of the alpha-amylase family predicted a (beta/alpha)8-barrel domain. Six of the eight highly conserved regions in amylolytic enzymes are present in amylosucrase. Among them, four constitute the active site in alpha-amylases. These sites were also conserved in the sequence of glucosyltransferases and dextransucrases. Nevertheless, the evolutionary tree does not show strong homology between them. The amylosucrase was purified by affinity chromatography between fusion protein glutathione S-transferase-amylosucrase and glutathione-Sepharose 4B. The pure enzyme linearly elongated some branched chains of glycogen, to an average degree of polymerization of 75.

Please choose payment method:






(PDF emailed within 1 workday: $29.90)

Accession: 003557667

Download citation: RISBibTeXText

PMID: 9882648


Related references

Cloning and characterization of the gene for amylosucrase from Neisseria polysaccharea: production of a linear a-1,4-glucan. Journal of Bacteriology 179(10): 24-30, 1997

Cloning and characterization of the gene for amylosucrase from Neisseria polysaccharea: production of a linear alpha-1,4-glucan. Journal of Bacteriology 179(10): 3324-3330, 1997

Crystallization and preliminary X-ray studies of recombinant amylosucrase from Neisseria polysaccharea. Acta Crystallographica Section D Biological Crystallography 56(2): 203-205, 2000

Amylosucrase from Neisseria polysaccharea: novel catalytic properties. Febs Letters 471(2-3): 219-223, 2000

Characterisation of the activator effect of glycogen on amylosucrase from Neisseria polysaccharea. FEMS Microbiology Letters 186(1): 103-108, 2000

Identification of key amino acid residues in Neisseria polysaccharea amylosucrase. FEBS Letters 474(1): 33-37, 2000

Crystal structure of the covalent intermediate of amylosucrase from Neisseria polysaccharea. Biochemistry 43(11): 3104-3110, 2004

Maltooligosaccharide disproportionation reaction: an intrinsic property of amylosucrase from Neisseria polysaccharea. Febs Letters 527(1-3): 67-70, 2002

Elongation and insolubilisation of alpha-glucans by the action of Neisseria polysaccharea amylosucrase. Journal of Cereal Science 40(1): 17-30, 2004

Molecular basis of the amylose-like polymer formation catalyzed by Neisseria polysaccharea amylosucrase. Journal of Biological Chemistry 279(1): 726-734, 2004

Engineering of anp efficient mutant of Neisseria polysaccharea amylosucrase for the synthesis of controlled size maltooligosaccharides. Carbohydrate Polymers 173: 403-411, 2017

Essential role of amino acid position 226 in oligosaccharide elongation by amylosucrase from Neisseria polysaccharea. Biotechnology and Bioengineering 111(9): 1719-1728, 2014

Crystal structure of the Glu328Gln mutant of Neisseria polysaccharea amylosucrase in complex with sucrose and maltoheptaose. Biocatalysis and Biotransformation 24(1-2): 99-105, 2006

Probing impact of active site residue mutations on stability and activity of Neisseria polysaccharea amylosucrase. Protein Science 22(12): 1754-1765, 2013

Synthesis of aesculetin and aesculin glycosides using engineered Escherichia coli expressing Neisseria polysaccharea amylosucrase. Journal of Microbiology and Biotechnology 2018:, 2018